Vinpocetine and eburn amonine derivatives for promoting bone growth

ABSTRACT

The present invention provides a method of promoting bone growth in a subject in need thereof, by administering to the subject a therapeutically effective amount of a compound of Formula I. The compounds include the salts, hydrates and isomers thereof. The present invention also provides methods for the treatment of renal disease and cancer.

CROSS-REFERENCES TO RELATED APPLICATIONS

This application claims priority to U.S. Provisional Application No.61/078,163, filed Jul. 3, 2008, which is incorporated in its entiretyherein for all purposes.

BACKGROUND OF THE INVENTION

It is well-understood that bone formation is indicated for treatment ofa wide variety of disparate disorders in mammals including simple aging,bone degeneration and osteoporosis, fracture healing, fusion orarthrodesis, osteogenesis imperfecta, etc., as well as for successfulinstallation of various medical orthopedic and periodontal implants suchas screws, rods, titanium cage for spinal fusion, hip joints, kneejoint, ankle joints, shoulder joints, dental plates and rods, etc.

Increasing bone mineralization to treat conditions characterized atleast in part by increased bone resorption, such as osteopenia, bonefractures, osteoporosis, arthritis, tumor metastases, Paget's diseaseand other metabolic bone disorders, using cathepsin K inhibitors andTGF-beta binding proteins, etc., are well-known as shown by U.S.Publication No. 2004/0235728 to Selwyn Aubrey Stoch, published Nov. 25,2004, and Mary E. Brunkow et al., U.S. Pat. No. 6,489,445 and U.S.Publication No. 2004/0009535, published Jan. 15, 2004. In the Brunkow'535 publication and '445 patent, the TGF-beta binding proteins includeSost polypeptide (full length and short peptide) antibodies thatinterfere with the interaction between the TGF-beta binding proteinsclerostin and a TGF-beta superfamily member, particularly a bonemorphogenic protein. All of the diseases named above are due to asystemic loss of bone mineral and thus the administration of theantibody therapeutic is for systemic (whole body) increase in bonemineral density.

In the Brunkow '535 publication and '445 patent, the binding proteinspreferably bind specifically to at least one human bone morphogenicprotein (BMP) among BMP-5 and BMP-6.

U.S. Pat. No. 6,395,511 to Brunkow, et al. teaches a novel family ofhuman TGF-beta binding proteins and nucleic acids encoding them. Theprotein binds to at least human bone morphogenic protein-5 and humanbone morphogenic protein-6.

Sclerosteosis is a progressive sclerosing bone dysplasia. Sclerostin(the Sost gene) was originally identified as the sclerosteosis-causinggene. Sclerostin was intensely expressed in developing bones of mouseembryos. Punctuated expression of sclerostin was localized on thesurfaces of both intramembranously forming skull bones andendochondrally forming long bones. The physiological role of sclerostinremains to be elucidated. However, it is known that loss of functionmutations in Sost cause a rare bone dysplasia characterized by skeletalovergrowth.

In U.S. Publication No. 2006/0165799, published Jul. 27, 2006, teaches abone-filling composition for stimulating bone-forming andbone-consolidation comprising biocompatible calcium sulfate and viscousbiopolymers. The composition is intended to be administered easily intothe missing part of injured bone without diffusing to surroundingorgans.

In U.S. Pat. No. 5,939,039, issued in 1999 teaches the processes toyield unique calcium phosphate precursor minerals that can be used toform a self-setting cement or paste. Once placed in the body, thesecalcium phosphate cements (CPC) will be resorbed and remodeled(converted) to bone.

For example, calcium phosphate particles prepared in accordance with the'039 patent can be used in any of the orthopedic or dental proceduresknown for the use of calcium phosphate; the procedures of bone fillingdefect repair, oncological defect filling, craniomaxillofacial voidfilling and reconstruction, dental extraction site filling.

U.S. Publication No. 2006/0198863 to Carl Alexander DePaula, publishedSep. 7, 2006, relates to a formable ceramic composition for filling bonedefects. The composition comprises ceramic beta tricalcium phosphateparticles having a particle size from about 40 microns to 500 micronsadmixed with a hydrogel carrier containing citric acid buffer. Thecomposition has a pH between 7.0 to 7.8 and the hydrogel component ofthe carrier ranges from about 1.0 to 5.0% of the composition.

Wise and SOST are understood to be closely related family members(Ellies et al., JBMR 2006 November; 21(11):1738-49.). Those of ordinaryskill are aware that the Wise null mutant mouse exhibits a bonephenotype (Keynote presentation at the 2005 American Society of BoneMineral Research meeting in Nashville, Tenn. State of the Art lectures,an embryonic source of skeletal tissue. Patterning CraniofacialDevelopment; by Robb Krumlauf, Ph.D., Stowers Institute for MedicalResearch, Kansas City, Mo., USA).

U.S. Publication No. 2005/025604 to Vignery published Nov. 17, 2005shows induction of bone formation by mechanically inducing an increasein osteoblast activity and elevating systemic blood concentration of abone anabolic agent, including optionally elevating systemic bloodconcentration of an antiresorptive agent.

Finally, Yanagita, Modulator of bone morphogenic protein activity in theprogression of kidney diseases, Kidney Int., Vol. 70, No. 6 (2006)989-93 shows Usag-1 (also known as “Wise”) protects the kidney fromcisplatin insult due to BMP inhibition. See also Yanagita, Uterinesensitization-associated gene-1 (USAG-1), a novel antagonist expressedin the kidney, accelerates tubular injury, J. Clin. Invest., Vol. 116,No. 1 (2005) 70-9, Yanagita, BMP antagonists: their roles in developmentand involvement in pathophysiology, Cytokine Growth Factor Rev., Vol 16,No. 3 (2005) 309-17, and Yanagita, USAG-1: a bone morphogenic proteinantagonist abundantly expressed in the kidney, Biochem. Biophys. Res.Commun., Vol. 316, No. 2 (2004) 490-500.

What is needed in the art is a new method for treating the bonedisorders described above, as well as others. Surprisingly, the presentinvention meets these and other needs.

BRIEF SUMMARY OF THE INVENTION

In one embodiment, the present invention provides a method of promotingbone growth in a subject in need thereof, comprising administering tothe subject a therapeutically effective amount of a compound of FormulaI:

wherein each of R¹, R², R⁵, R⁷ or R⁸ are independently H, halogen, C₁₋₆alkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₁₋₆haloalkoxy, C₁₋₆ alkyl-hydroxy, —OR^(1a), —C(O)R^(1a), —C(O)OR^(1a),—C(O)NR^(1a)R^(1b), —NR^(1a)R^(1b), —SR^(1a), —N(R^(1a))C(O)R^(1b),—N(R^(1a))C(O)OR^(1b), —N(R^(1a))C(O)NR^(1a)R^(1b), —OP(O)(OR^(1a))₂,—S(O)₂OR^(1a), —S(O)₂NR^(1a)R^(1b), —CN, —CH═N—OH, C₁₋₆alkyl-OC(O)-aryl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl.Alternatively, R¹ is combined with R² to form ═O or ═NR^(1a), R¹ iscombined with R³ to form a bond, or R¹ is combined with R⁵ to form acycloalkyl. Each R^(1a) or R^(1b) is H, C₁₋₆ alkyl, C₁₋₆ alkyl-hydroxy,C₁₋₆ alkyl-NO₂, C₁₋₆ alkyl-cycloalkyl, heterocycloalkyl, or heteroaryl.Each of R³ or R⁴ are H, C₁₋₆ alkyl or C₁₋₆ haloalkyl. Alternatively twoR⁵ or R⁷ groups attached to the same carbon can be combined to form ═O.R⁶ is absent or an N-oxide. The compounds include the salts, hydratesand isomers thereof. Accordingly, administration of the compound resultsin bone growth in the subject.

In a second embodiment, the present invention provides a method ofpromoting bone growth in a subject in need thereof, comprisingadministering to the subject a therapeutically effective amount of acompound of Formula I, thereby promoting bone growth in the subject.

In a third embodiment, the present invention provides a method oftreating renal damage, comprising administering to a subject in needthereof, a therapeutically effective amount of a compound of the presentinvention.

In a fourth embodiment, the present invention provides an orthopedic orperiodontal medical device comprising a structural support, wherein animplantable portion of the structural support is adapted to bepermanently implanted within a subject, wherein the implantable portionis attached to a bone, the structural support bearing at least a partialexternal coating comprising a compound of the present invention.

In a fifth embodiment, the present invention provides a method oftreating cancer, comprising administering to a subject in need thereof,a therapeutically effective amount of a compound of Formula I.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the increase in tibia long bone bone-volume/total volumeover baseline control: 17% at 5 mg/kg (B) and 10% at 1 mg/kg (C) ofeburnamonine phosphate in mice.

FIG. 2 shows the percent change in serum osteocalcin increase versussaline controls of 35% at 5 mg/kg of eburnamonine phosphate in mice.

FIG. 3 shows the increase in tibia long bone bone-volume/total volumewhen compared to baseline controls: 9% at 5 mg/kg (D) and 17% at 0.33mg/kg (E) of eburnamonine phosphate in mice.

FIG. 4 shows the increase in osteocalcin serum levels, of 62% and 39%,for vinpocetine citrate at 10 mg/kg and ebumamonine phosphate at 0.33mg/kg, respectively.

DETAILED DESCRIPTION OF THE INVENTION I. General

The present invention encompasses compounds, compositions and methodsfor promoting bone growth in a subject. The compounds of the presentinvention are SOST (Sclerostin) and Wise antagonists that modulate theWnt pathway. By modulating the Wnt pathway, the compounds andcompositions of the present invention promote bone growth. The bonegrowth can be systemic or local bone growth. The compounds andcompositions of the present invention can be administered locally orsystemically. The present invention also provides implantable devicesfor delivering the compounds and compositions of the present invention.The compounds and compositions of the present invention also act totreat renal damage and cancer.

II. Definitions

As used herein, the term “pharmaceutically acceptable excipient” refersto a substance that aids the administration of an active agent to andabsorption by a subject. Pharmaceutically acceptable excipients usefulin the present invention include, but are not limited to, binders,fillers, disintegrants, lubricants, coatings, sweeteners, flavors andcolors. One of skill in the art will recognize that other pharmaceuticalexcipients are useful in the present invention.

As used herein, the term “alkyl” refers to a straight or branched,saturated, aliphatic radical having the number of carbon atomsindicated. For example, C₁-C₆ alkyl includes, but is not limited to,methyl, ethyl, propyl, butyl, pentyl, hexyl, iso-propyl, iso-butyl,sec-butyl, tert-butyl, etc.

Alkylene represents either straight chain or branched alkylene of 1 to 7carbon atoms, i.e. a divalent hydrocarbon radical of 1 to 7 carbonatoms; for instance, straight chain alkylene being the bivalent radicalof Formula —(CH₂)_(n″), where n is 1, 2, 3, 4, 5, 6 or 7. Preferablyalkylene represents straight chain alkylene of 1 to 4 carbon atoms, e.g.a methylene, ethylene, propylene or butylene chain, or the methylene,ethylene, propylene or butylene chain mono-substituted by C₁-C₃-alkyl(preferably methyl) or disubstituted on the same or different carbonatoms by C₁-C₃-alkyl (preferably methyl), the total number of carbonatoms being up to and including 7.

As used herein, the term “alkoxy” refers to alkyl with the inclusion ofan oxygen atom, for example, methoxy, ethoxy, etc.“Halo-substituted-alkoxy” is as defined for alkoxy where some or all ofthe hydrogen atoms are substituted with halogen atoms. For example,halo-substituted-alkoxy includes trifluoromethoxy, etc.

As used herein, the term “alkenyl” refers to either a straight chain orbranched hydrocarbon of 2 to 6 carbon atoms, having at least one doublebond. Examples of alkenyl groups include, but are not limited to, vinyl,propenyl, isopropenyl, butenyl, isobutenyl, butadienyl, pentenyl orhexadienyl.

As used herein, the term “alkynyl” refers to either a straight chain orbranched hydrocarbon of 2 to 6 carbon atoms, having at least one triplebond. Examples of alkynyl groups include, but are not limited to,acetylenyl, propynyl or butynyl.

As used herein, the term “halogen” refers to fluorine, chlorine, bromineand iodine.

As used herein, the term “haloalkyl” refers to alkyl as defined abovewhere some or all of the hydrogen atoms are substituted with halogenatoms. Halogen (halo) preferably represents chloro or fluoro, but mayalso be bromo or iodo. For example, haloalkyl includes trifluoromethyl,fluoromethyl, 1,2,3,4,5-pentafluoro-phenyl, etc. The term “perfluoro”defines a compound or radical which has at least two available hydrogenssubstituted with fluorine. For example, perfluorophenyl refers to1,2,3,4,5-pentafluorophenyl, perfluoromethane refers to1,1,1-trifluoromethyl, and perfluoromethoxy refers to1,1,1-trifluoromethoxy.

As used herein, the term “cycloalkyl” refers to a saturated or partiallyunsaturated, monocyclic, fused bicyclic or bridged polycyclic ringassembly containing from 3 to 12 ring atoms, or the number of atomsindicated For example, C₃₋₈cycloalkyl includes cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, and up to cyclooctyl.

As used herein, the term “heterocycle” refers to a ring system havingfrom 3 ring members to about 20 ring members and from 1 to about 5heteroatoms such as N, O and S. Additional heteroatoms can also beuseful, including, but not limited to, B, Al, Si and P. The heteroatomscan also be oxidized, such as, but not limited to, —S(O)— and —S(O)₂—.For example, heterocycle includes, but is not limited to,tetrahydrofuranyl, tetrahydrothiophenyl, morpholino, pyrrolidinyl,pyrrolinyl, imidazolidinyl, imidazolinyl, pyrazolidinyl, pyrazolinyl,piperazinyl, piperidinyl, indolinyl, quinuclidinyl and1,4-dioxa-8-aza-spiro[4.5]dec-8-yl.

As used herein, the term “aryl” refers to a monocyclic or fusedbicyclic, tricyclic or greater, aromatic ring assembly containing 6 to16 ring carbon atoms. For example, aryl may be phenyl, benzyl ornaphthyl, preferably phenyl. “Arylene” means a divalent radical derivedfrom an aryl group. Aryl groups can be mono-, di- or tri-substituted byone, two or three radicals selected from alkyl, alkoxy, aryl, hydroxy,halogen, cyano, amino, amino-alkyl, trifluoromethyl, alkylenedioxy andoxy-C₂-C₃-alkylene; all of which are optionally further substituted, forinstance as hereinbefore defined; or 1- or 2-naphthyl; or 1- or2-phenanthrenyl. Alkylenedioxy is a divalent substitute attached to twoadjacent carbon atoms of phenyl, e.g. methylenedioxy or ethylenedioxy.Oxy-C₂-C₃-alkylene is also a divalent substituent attached to twoadjacent carbon atoms of phenyl, e.g. oxyethylene or oxypropylene. Anexample for oxy-C₂-C₃-alkylene-phenyl is 2,3-dihydrobenzofuran-5-yl.

Preferred as aryl is naphthyl, phenyl or phenyl mono- or disubstitutedby alkoxy, phenyl, halogen, alkyl or trifluoromethyl, especially phenylor phenyl-mono- or disubstituted by alkoxy, halogen or trifluoromethyl,and in particular phenyl.

Examples of substituted phenyl groups as R are, e.g. 4-chlorophen-1-yl,3,4-dichlorophen-1-yl, 4-methoxyphen-1-yl, 4-methylphen-1-yl,4-aminomethylphen-1-yl, 4-methoxyethylaminomethylphen-1-yl,4-hydroxyethylaminomethylphen-1-yl,4-hydroxyethyl-(methyl)-aminomethylphen-1-yl, 3-aminomethylphen-1-yl,4-N-acetylaminomethylphen-1-yl, 4-aminophen-1-yl, 3-aminophen-1-yl,2-aminophen-1-yl, 4-phenyl-phen-1-yl, 4-(imidazol-1-yl)-phen-yl,4-(imidazol-1-ylmethyl)-phen-1-yl, 4-(morpholin-1-yl)-phen-1-yl,4-(morpholin-1-ylmethyl)-phen-1-yl,4-(2-methoxyethylaminomethyl)-phen-1-yl and4-(pyrrolidin-1-ylmethyl)-phen-1-yl, 4-(thiophenyl)-phen-1-yl,4-(3-thiophenyl)-phen-1-yl, 4-(4-methylpiperazin-1-yl)-phen-1-yl, and4-(piperidinyl)-phenyl and 4-(pyridinyl)-phenyl optionally substitutedin the heterocyclic ring.

Similarly, substituents for the aryl and heteroaryl groups are variedand are selected from: -halogen, —OR′, —OC(O)R′, —NR′R″, —SR″, —R′, —CN,—NO₂, —CO₂R′, —CONR′R″, —C(O)R′, —OC(O)NR′R″, —NR″C(O)R′, —NR″C(O)₂R′,—NR′—C(O)NR″R′″, —NH—C(NH₂)═NH, —NR′C(NH₂)═NH, —NH—C(NH₂)═NR′, —S(O)R′,—S(O)₂R′, —S(O)₂NR′R″, —N₃, —CH(Ph)₂, perfluoro(C₁-C₄)alkoxy, andperfluoro(C₁-C₄)alkyl, in a number ranging from zero to the total numberof open valences on the aromatic ring system; and where R′, R″ and R′″are independently selected from hydrogen, (C₁-C₈)alkyl and heteroalkyl,unsubstituted aryl and heteroaryl, (unsubstituted aryl)-(C₁-C₄)alkyl,and (unsubstituted aryl)oxy-(C₁-C₄)alkyl.

Two of the substituents on adjacent atoms of the aryl or heteroaryl ringmay optionally be replaced with a substituent of the formula-T-C(O)—(CH₂)_(q)—U—, wherein T and U are independently —NH—, —O—, —CH₂—or a single bond, and q is an integer of from 0 to 2. Alternatively, twoof the substituents on adjacent atoms of the aryl or heteroaryl ring mayoptionally be replaced with a substituent of the formula-A-(CH₂)_(r)—B—, wherein A and B are independently —CH₂—, —O—, —NH—,—S—, —S(O)—, —S(O)₂—, —S(O)₂NR′— or a single bond, and r is an integerof from 1 to 3. One of the single bonds of the new ring so formed mayoptionally be replaced with a double bond. Alternatively, two of thesubstituents on adjacent atoms of the aryl or heteroaryl ring mayoptionally be replaced with a substituent of the formula—(CH₂)_(s)—X—(CH₂)_(t)—, where s and t are independently integers offrom 0 to 3, and X is —O—, —NR′—, —S—, —S(O)—, —S(O)₂—, or —S(O)₂NR′—.The substituent R′ in —NR′— and —S(O)₂NR′— is selected from hydrogen orunsubstituted (C₁-C₆)alkyl.

As used herein, the term “heteroaryl” refers to a monocyclic or fusedbicyclic or tricyclic aromatic ring assembly containing 5 to 16 ringatoms, where from 1 to 4 of the ring atoms are a heteroatom each N, O orS. For example, heteroaryl includes pyridyl, indolyl, indazolyl,quinoxalinyl, quinolinyl, isoquinolinyl, benzothienyl, benzofuranyl,furanyl, pyrrolyl, thiazolyl, benzothiazolyl, oxazolyl, isoxazolyl,triazolyl, tetrazolyl, pyrazolyl, imidazolyl, thienyl, or any otherradicals substituted, especially mono- or di-substituted, by e.g. alkyl,nitro or halogen. Pyridyl represents 2-, 3- or 4-pyridyl, advantageously2- or 3-pyridyl. Thienyl represents 2- or 3-thienyl. Quinolinylrepresents preferably 2-, 3- or 4-quinolinyl. Isoquinolinyl representspreferably 1-, 3- or 4-isoquinolinyl. Benzopyranyl, benzothiopyranylrepresents preferably 3-benzopyranyl or 3-benzothiopyranyl,respectively. Thiazolyl represents preferably 2- or 4-thiazolyl, andmost preferred, 4-thiazolyl. Triazolyl is preferably 1-, 2- or5-(1,2,4-triazolyl). Tetrazolyl is preferably 5-tetrazolyl.

Preferably, heteroaryl is pyridyl, indolyl, quinolinyl, pyrrolyl,thiazolyl, isoxazolyl, triazolyl, tetrazolyl, pyrazolyl, imidazolyl,thienyl, furanyl, benzothiazolyl, benzofuranyl, isoquinolinyl,benzothienyl, oxazolyl, indazolyl, or any of the radicals substituted,especially mono- or di-substituted.

Similarly, substituents for the aryl and heteroaryl groups are variedand are selected from: -halogen, —OR′, —OC(O)R′, —NR′R″, —SR′, —R′, —CN,—NO₂, —CO₂R′, —CONR′R″, —C(O)R′, —OC(O)NR′R″, —NR″C(O)R′, —NR″C(O)₂R′,—NR′—C(O)NR″R′″, —NH—C(NH₂)═NH, —NR′C(NH₂)═NH, —NH—C(NH₂)═NR′, —S(O)R′,—S(O)₂R′, —S(O)₂NR′R″, —N₃, —CH(Ph)₂, perfluoro(C₁-C₄)alkoxy, andperfluoro(C₁-C₄)alkyl, in a number ranging from zero to the total numberof open valences on the aromatic ring system; and where R′, R″ and R′″are independently selected from hydrogen, (C₁-C₈)alkyl and heteroalkyl,unsubstituted aryl and heteroaryl, (unsubstituted aryl)-(C₁-C₄)alkyl,and (unsubstituted aryl)oxy-(C₁-C₄)alkyl.

Two of the substituents on adjacent atoms of the aryl or heteroaryl ringmay optionally be replaced with a substituent of the formula-T-C(O)—(CH₂)_(q)—U—, wherein T and U are independently —NH—, —O—, —CH₂—or a single bond, and q is an integer of from 0 to 2. Alternatively, twoof the substituents on adjacent atoms of the aryl or heteroaryl ring mayoptionally be replaced with a substituent of the formula-A-(CH₂)_(r)—B—, wherein A and B are independently —CH₂—, —O—, —NH—,—S—, —S(O)—, —S(O)₂—, —S(O)₂NR′— or a single bond, and r is an integerof from 1 to 3. One of the single bonds of the new ring so formed mayoptionally be replaced with a double bond. Alternatively, two of thesubstituents on adjacent atoms of the aryl or heteroaryl ring mayoptionally be replaced with a substituent of the formula—(CH₂)_(s)—X—(CH₂)_(t)—, where s and t are independently integers offrom 0 to 3, and X is —O—, —NR′—, —S—, —S(O)—, —S(O)₂—, or —S(O)₂NR′—.The substituent R′ in —NR′— and —S(O)₂NR′— is selected from hydrogen orunsubstituted (C₁-C₆)alkyl.

As used herein, the term “salt” refers to acid or base salts of thecompounds used in the methods of the present invention. Illustrativeexamples of pharmaceutically acceptable salts are mineral acid(hydrochloric acid, hydrobromic acid, phosphoric acid, phosphonic acidand the like) salts, organic acid (acetic acid, propionic acid, glutamicacid, citric acid and the like) salts, quaternary ammonium (methyliodide, ethyl iodide, and the like) salts. It is understood that thepharmaceutically acceptable salts are non-toxic. Additional informationon suitable pharmaceutically acceptable salts can be found inRemington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company,Easton, Pa., 1985, which is incorporated herein by reference.

Pharmaceutically acceptable salts of the acidic compounds of the presentinvention are salts formed with bases, namely cationic salts such asalkali and alkaline earth metal salts, such as sodium, lithium,potassium, calcium, magnesium, as well as ammonium salts, such asammonium, trimethyl-ammonium, diethylammonium, andtris-(hydroxymethyl)-methyl-ammonium salts. Other salts include, but arenot limited to, citrate and phosphonic acid.

Similarly acid addition salts, such as of mineral acids, organiccarboxylic and organic sulfonic acids, e.g., hydrochloric acid,methanesulfonic acid, maleic acid, are also possible provided a basicgroup, such as pyridyl, constitutes part of the structure.

The neutral forms of the compounds can be regenerated by contacting thesalt with a base or acid and isolating the parent compound in theconventional manner. The parent form of the compound differs from thevarious salt forms in certain physical properties, such as solubility inpolar solvents, but otherwise the salts are equivalent to the parentform of the compound for the purposes of the present invention.

As used herein, the term “hydrate” refers to a compound that iscomplexed to at least one water molecule. The compounds of the presentinvention can be complexed with from 1 to 10 water molecules.

Certain compounds of the present invention possess asymmetric carbonatoms (optical centers) or double bonds; the racemates, diastereomers,geometric isomers and individual isomers are all intended to beencompassed within the scope of the present invention.

As used herein, the term “subject” refers to animals such as mammals,including, but not limited to, primates (e.g., humans), cows, sheep,goats, horses, dogs, cats, rabbits, rats, mice and the like. In certainembodiments, the subject is a human.

As used herein, the terms “therapeutically effective amount or dose” or“therapeutically sufficient amount or dose” or “effective or sufficientamount or dose” refer to a dose that produces therapeutic effects forwhich it is administered. The exact dose will depend on the purpose ofthe treatment, and will be ascertainable by one skilled in the art usingknown techniques (see, e.g., Lieberman, Pharmaceutical Dosage Forms(vols. 1-3, 1992); Lloyd, The Art, Science and Technology ofPharmaceutical Compounding (1999); Pickar, Dosage Calculations (1999);and Remington: The Science and Practice of Pharmacy, 20th Edition, 2003,Gennaro, Ed., Lippincott, Williams & Wilkins). In sensitized cells, thetherapeutically effective dose can often be lower than the conventionaltherapeutically effective dose for non-sensitized cells.

As used herein, the term “calcium salt” refers to salts containingcalcium. Examples of calcium salts include, but are not limited to,calcium acetate, calcium aluminates, calcium aluminosilicate, calciumarsenate, calcium borate, calcium bromide, calcium carbide, calciumcarbonate, calcium chlorate, calcium chloride, calcium citrate, calciumcitrate malate, calcium cyanamide, calcium dihydrogen phosphate, calciumfluoride, calcium formate, calcium glubionate, calcium glucoheptonate,calcium gluconate, calcium glycerylphosphate, calcium hexaboride,calcium hydride, calcium hydroxide, calcium hypochlorite, calciuminosinate, calcium iodate, calcium iodide, calcium lactate, calciumlactate gluconate, calcium magnesium acetate, calcium malate, calciumnitrate, calcium nitride, calcium oxalate, calcium oxide, calciumpangamate, calcium peroxide, calcium phosphate, calcium phosphide,calcium propionate, calcium pyrophosphate, calcium silicate, calciumsilicide, calcium sorbate, calcium stearate, calcium sulfate, calciumsulfide, calcium tartrate, calcium(I) chloride, dicalcium citrate,dicalcium phosphate, dodecacalcium hepta-aluminate, tricalciumaluminate, tricalcium phosphate and triple superphosphate. One of skillin the art will appreciate that other calcium salts are useful in thepresent invention.

As used herein, the term “site of injury or localized condition” refersto a specific location in the subject's body that is in need oftreatment by the method of the present invention. For example, theinjury can be a fracture and the localized condition can be a diseasestate (such as osteoporosis, etc.) that is limited to a particularlocation in the subject's body, such as a particular bone, joint, digit,hand, foot, limb, spine, head, torso, etc.

As used herein, the term “promoting bone growth” refers to thestimulation of new bone growth, or an increase in bone density or bonemineral content.

As used herein, the term “arthrodesis” refers to the artificialinduction of joint ossification between two bones, often via surgery.Arthrodesis can be accomplished via bone graft, metal implants or theuse of synthetic bone substitutes, among others.

As used herein, the term “bone autograft” refers to the grafting of asubject's own bone.

As used herein, the term “bone allograft” refers to the grafting of bonefrom one person to another person.

As used herein, the term “antiresorptive drug” refers to drugs that slowor block the resorption of bone.

As used herein, the term “bone related disease characterized by low bonemass” refers to bone having a T-score less than −1. Other methods ofdetermining low bone mass are known by one of skill in the art.

As used herein, the term “bone fracture” refers to bone that has beencracked or broken.

As used herein, the term “spinal fusion” refers to a surgical techniquefor combining two or more vertebrae.

As used herein, the term “structural support” refers to a segment of thedevice that can be implanted in a subject (implantable portion). Thestructural support can be prepared from a variety of differentmaterials, including metals, ceramics, polymers and inorganic materials,such as described below. The structural support can be coated with avariety of materials that promote bone growth.

As used herein, the term “external coating” refers to a coating of thestructural support that can cover only a portion of the structuralsupport (partial external coating) or cover the entire structuralsupport. For example, the partial external coating can completely coveronly the implantable portion of the structural support.

As used herein, the term “weakened bone” refers to bone that has a Tscore of less than −0.5 (less than 0.9 g/cm2).

As used herein, the term “demineralized bone” refers to bone from whichcalcium and phosphate have been removed. The remaining material containsthe osteoinductive proteins contained in the matrix. These proteinsinclude bone morphogenetic proteins that induce new bone formation.Demineralized bone often comes in the form of “demineralized bone matrix(DBM).” DBM can be made by fresh frozen or freeze-dried bulk boneallograft, or can be made from mild acid extraction of cadaveric bonethat removes the mineral phase, leaving collagen, growth factors, andnoncollagenous proteins that offer the intrinsic properties ofosteoconduction. DBM can also be processed in a variety of ways,ultimately resulting in a powder that is mixed with a carrier to providethe optimum handling characteristics desired by a surgeon. DBM isclinically available in gels, pastes, putty, and fabrics that have beentailored to meet the needs of the surgical procedure. Some DBM are mixedwith antibiotics prior to the surgical procedure.

As used herein, the term “renal damage” refers to the inability of thekidneys to excrete waste and to help maintain the electrolyte balance ofthe body. Renal damage is characterized by some of the following: highblood pressure, accumulation of urea and formation of uremic frost,accumulation of potassium in the blood, decrease in erythropoietinsynthesis, increase in fluid volume, hyperphosphatemia, and metabolicacidosis, among others.

As used herein, the term “osteoconductive matrix” refers to a materialthat can act as an osteoconductive substrate and has a scaffoldingstructure on which infiltrating cells can attach, proliferate, andparticipate in the process of producing osteoid, the organic phase ofbone, culminating in osteoneogenesis, or new bone formation. Matrix orscaffold means the structural component or substrate intrinsicallyhaving a 3 dimensional form upon which the specific cellular eventsinvolved in bone formation will occur. The osteoconductive matrix allowsfor the ingrowth of host capillaries, perivascular tissue andosteoprogenitor cells. The osteoconductive matrix can also include anosteoinductive agent for providing osteogenic potential. Anosteoinductive agent stimulates the host to build new bone.

As used herein, the terms “treat”, “treating” and “treatment” refers toany indicia of success in the treatment or amelioration of an injury,pathology, condition, or symptom (e.g., pain), including any objectiveor subjective parameter such as abatement; remission; diminishing ofsymptoms or making the symptom, injury, pathology or condition moretolerable to the patient; decreasing the frequency or duration of thesymptom or condition; or, in some situations, preventing the onset ofthe symptom or condition. The treatment or amelioration of symptoms canbe based on any objective or subjective parameter; including, e.g., theresult of a physical examination.

As used herein, the term “RankL inhibitor” refers to compounds or agentsthat inhibit the activity of RankL. RankL (Receptor Activator forNuclear Factor κ B Ligand), is important in bone metabolism byactivating osteoclasts. RankL inhibitors include, but are not limitedto, the human monoclonal antibody denosumab. One of skill in the artwill appreciate that other RankL inhibitors are useful in the presentinvention.

III. Compounds

The compounds useful in the methods of the present invention include anyvinpocetine and eburnamonine derivative. In some embodiments, thecompounds of the present invention have Formula I:

wherein each of R¹, R², R⁵, R⁷ or R⁸ are independently H, halogen, C₁₋₆alkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₁₋₆haloalkoxy, C₁₋₆ alkyl-hydroxy, —OR^(1a) —C(O)R^(1a), —C(O)OR^(1a),—C(O)NR^(1a)R^(1b), —NR^(1a)R^(1b), —SR^(1a), —N(R^(1a))C(O)R^(1b),—N(R^(1a))C(O)OR^(1b), —N(R^(1a))C(O)NR^(1a)R^(1b), —OP(O)(OR^(1a))₂,—S(O)₂OR^(1a), —S(O)₂NR^(1a)R^(1b), —CN, —CH═N—OH, C₁₋₆alkyl-OC(O)-aryl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl.Alternatively, R¹ is combined with R² to form ═O or ═NR^(1a), R¹ iscombined with R³ to form a bond, or R¹ is combined with R⁵ to form acycloalkyl. Each R^(1a) or R^(1b) is H, C₁₋₆ alkyl, C₁₋₆ alkyl-hydroxy,C₁₋₆ alkyl-NO₂, C₁₋₆ alkyl-cycloalkyl, heterocycloalkyl, or heteroaryl.Each of R³ or R⁴ are H, C₁₋₆ alkyl or C₁₋₆ haloalkyl. Alternatively twoR⁵ or R⁷ groups attached to the same carbon can be combined to form ═O.R⁶ is absent or an N-oxide.

In other embodiments, the compound has Formula Ia:

In some other embodiments, the compound has the formula:

In still other embodiments, R^(1a) is C₁₋₆ alkyl and R⁴ is C₁₋₆ alkyl.In yet other embodiments, the compound has the formula:

In other embodiments, the compound has Formula Ib:

In some other embodiments, the compound has the formula:

Compounds of Formula I useful in the methods of the present inventionare described in the table below.

TABLE I Compounds of Formula I. (I)

Compound R¹ R² R³ R⁴ R⁵ R⁶ R⁷ R⁸ 1 C(O)OMe combined to Et H — H H form abond 2 C(O)OH combined to Et H — H H form a bond 3 C(O)OEt combined toEt H — H H form a bond 4 C(O)OEt combined to Et H — H H form a bond 5C(O)OMe combined to Et H N- H H form a bond oxide 6

combined to form a bond Et H — H H 7

combined to form a bond Et H — H H 8 CH═NOH combined to Et H — H H forma bond 9 C(O)OMe combined to Et H — 6- H form a bond (═O) 10 C(O)OMecombined to Et H — H 11-Br form a bond 11 C(O)OCH₂OCH₃ combined to Et H— H H form a bond 12

combined to form a bond Et H — H H 13

combined to form a bond Et H — H H 14 H combined to H H — H 11- form abond Me 15 C(O)OCH₂CH₂ONO₂ combined to Et H — H H form a bond 16 C(O)OEtcombined to CH₂CF₃ H — H H form a bond 17 H OH H Et H — H H 18 ═O H Et H— H H 19 C(O)OMe OH H Et H — H H 20 CH₂OH OH H Et H — H H 21 C(O)OMe H HEt H — H H 22 C(O)OH OH H Et H — H H 23 C(O)OEt OH H Et H — H H 24C(O)OMe OH H Et H N- H H oxide 25 C(O)OEt OH H Et H N- H H oxide 26C(O)OMe OH H 16,17- — H 12- ethyleneoxy OMe 27 C(O)OMe OH H Et H N- H11-Br oxide 28 C(O)OCH₂CH(OH)CH₃ H H Et H — H H 29 C(O)NH₂ H H Et H — HH 30

H H Et H — H H 31 CH═NOH H H Et H — H H 32 ═O H Et H — 6- H (═0) 33C(O)OMe OH H Et H — 6- H (═O) 34 C(O)OMe OH H Et H — 6-OH H 35 CH₂OH OHH Et H — 6- H (═O) 36 H OH H H H — H H 37 ═NH H Et H — H H 38 C(O)OMe OHH Et H H H 11-Br 39 C(O)OH OH H Et H H H 11-Br 40 H H H H H — H H 41 HOH H H H — H H 42 ═O H Et H — 6-OH H 43 CH₂OH H H Et H — H H

The compounds of the present invention also include the salts, hydrates,solvates and prodrug forms. The compounds of the present invention alsoinclude the isomers and metabolites of those described in Formula I.

The compounds of the present invention can be in the salt form. Saltsinclude, but are not limited, to sulfate, citrate, acetate, oxalate,chloride, bromide, iodide, nitrate, bisulfate, phosphate, acidphosphate, phosphonic acid, isonicotinate, lactate, salicylate, citrate,tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate,succinate, maleate, gentisinate, fumarate, gluconate, glucaronate,saccharate, formate, benzoate, glutamate, methanesulfonate,ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamoate(i.e., 1,1′-methylene-bis-(2-hydroxy-3-naphthoate)) salts. Other saltsinclude, but are not limited to, salts with inorganic bases includealkali metal salts such as sodium salts, and potassium salts; alkalineearth metal salts such as calcium salts, and magnesium salts; aluminumsalts; and ammonium salts. Other salts with organic bases include saltswith diethylamine, diethanolamine, meglumine, andN,N′-dibenzylethylenediamine.

The compounds of the present invention can be made by a variety ofmethods known to one of skill in the art (see Comprehensive OrganicTransformations Richard C. Larock, 1989) One of skill in the art willappreciate that other methods of making the compounds are useful in thepresent invention.

IV. Administration

In some embodiments, the present invention provides a pharmaceuticalcomposition including a pharmaceutically acceptable excipient and acompound of Formula I. In other embodiments, the composition furthercomprises an osteoconductive matrix.

The compounds and compositions of the present invention can beadministered locally or systemically.

A. Local Delivery

It also has been found that successful implantation of the compounds ofthe present invention for bone formation requires association of thecompounds with a suitable carrier material capable of maintaining thecompound at an in vivo site of application. The carrier can bebiocompatible, a matrix, in vivo biodegradable and porous enough toallow cell infiltration.

The Sost or Wise antagonists are useful in clinical applications inconjunction with a suitable delivery or support system (matrix). Asdisclosed herein, the matrix can be combined with Sost or Wiseantagonist to induce bone formation reliably and reproducibly in amammalian body. The matrix preferably includes particles of porousmaterials. The pores are preferred to be of a dimension to permitprogenitor cell migration into the matrix and subsequent differentiationand proliferation. The particle size can be within the range of 70um-850 um, preferably 70 um-420 um, most preferably 150 um-420 um. Itcan be fabricated by close packing particulate material into a shapespanning the bone defect, or by otherwise structuring as desired amaterial that is biocompatible, and preferably biodegradable in vivo toserve as a “temporary scaffold” and substratum for recruitment ofmigratory progenitor cells, and as a base for their subsequent anchoringand proliferation.

In some embodiments, the matrix can be an osteoconducive matrix. Theosteoconducive matrix can include an osteoinducive agent and,optionally, a structural support. The osteoinductive agent can be anyagent that promotes bone formation. In some embodiments, theosteoinductive agent can be bone allograft, bone autograft,demineralized bone or periodontal ligament cells. The osteoconductivematrix can also include a structural support such as a calcium salt,calcium sulfate, calcium phosphate, a calcium phosphate cement,hydroxyapatite, coralline based hydroyxapatite (HA), dicalciumphosphate, tricalcium phosphate (TCP), calcium carbonate, collagen,plaster of Paris, phosphosphoryn, a borosilicate, a biocompatibleceramic, a calcium phosphate ceramic and polytetrafluoroethylene.

Other useful matrix materials include, for example, collagen;homopolymers or copolymers of glycolic acid, lactic acid, and butyricacid, including derivatives thereof, and ceramics, hydroxyapatite,tricalcium phosphate and other calcium phosphates, and calciumsulphates. Other matrices useful in the present invention include, butare not limited to, Kryptonite bone cement (Doctors Research Group,Oxford, Conn.) and Genex bone graft (Biocomposites, Wilmington, N.C.).Combinations of these matrix materials also can be useful.

When the SOST antagonist candidate is delivered in a carrier, thecontrol solution is ideally the carrier absent the SOST antagonistcandidate. Multiple doses of the SOST antagonist candidate can beapplied to the test animal, preferably following a predeterminedschedule of dosing. The dosing schedule can be over a period of days,more preferably over a period of weeks.

B. Systemic Delivery

In an exemplary embodiment, localized injection in situ of a SOSTantagonist candidate, can be made into a test animal, with a controlanimal receiving an equal volume of control solution without the SOSTantagonist candidate. Suitable dosage will depend on the nature of theparticular SOST antagonist candidate being tested. By way of example, indosing it should be noted that systemic injection, either intravenously,subcutaneously or intramuscularly, can also be used. Dosing performed bynebulized inhalation, eye drops, or oral ingestion should be at anamount sufficient to produce blood levels of the SOST antagonistcandidate similar to those reached using systemic injection. The amountof SOST antagonist candidate that must be delivered by nebulizedinhalation, eye drops, or oral ingestion to attain these levels isdependent upon the nature of the inhibitor used and can be determined byroutine experimentation.

Individuals to be treated using methods of the present invention can beany mammal, for example local increase in bone can be used for fracturehealing, fusion (arthrodesis), orthopedic reconstruction, andperiodontal repair. Systemic increase in bone would be for treatment oflow bone mass, i.e. osteoporosis. Such individuals include a dog, cat,horse, cow, or goat, particularly a commercially important animal or adomesticated animal, more particularly a human.

In therapeutic use SOST antagonists generally will be in the form of apharmaceutical composition containing the antagonist and apharmaceutically acceptable carrier. Pharmaceutically acceptablecarriers are well known in the art and include aqueous solutions such asphysiologically buffered saline or other buffers or solvents or vehiclessuch as glycols, glycerol, oils such as olive oil or injectable organicesters. The selection of a pharmaceutically acceptable carrier willdepend, in part, on the chemical nature of the SOST antagonist.

A pharmaceutically acceptable carrier may include physiologicallyacceptable compounds that act, for example, to stabilize the SOSTantagonist or increase its absorption, or other excipients as desired.Physiologically acceptable compounds include, for example,carbohydrates, such as glucose, sucrose or dextrans, antioxidants, suchas ascorbic acid or glutathione, chelating agents, low molecular weightproteins or other stabilizers or excipients. One skilled in the artwould know that the choice of a pharmaceutically acceptable carrier,including a physiologically acceptable compound, depends, for example,on the route of administration of the SOST antagonist and on itsparticular physio-chemical characteristics.

Generally, such carriers should be nontoxic to recipients at the dosagesand concentrations employed. Ordinarily, the preparation of suchcompositions entails combining the therapeutic agent with buffers,antioxidants such as ascorbic acid, low molecular weight (less thanabout 10 residues) polypeptides, proteins, amino acids, carbohydratesincluding glucose, maltose, sucrose or dextrins, chelating agents suchas EDTA, glutathione and other stabilizers and excipients. Neutralbuffered saline or saline mixed with nonspecific serum albumin areexemplary appropriate diluents.

The pharmaceutical compositions of the present invention can be preparedfor administration by a variety of different routes. In general, thetype of carrier is selected based on the mode of administration.Pharmaceutical compositions can be formulated for any appropriate mannerof administration, including, for example, topical, oral, nasal,intrathecal, rectal, vaginal, sublingual or parenteral administration,including subcutaneous, intravenous, intramuscular, intrasternal,intracavernous, intrameatal, or intraurethral injection or infusion. Apharmaceutical composition (e.g., for oral administration or delivery byinjection) can be in the form of a liquid (e.g., an elixir, syrup,solution, emulsion or suspension). A liquid pharmaceutical compositionmay include, for example, one or more of the following: sterile diluentssuch as water for injection, saline solution, preferably physiologicalsaline, Ringer's solution, isotonic sodium chloride, fixed oils that mayserve as the solvent or suspending medium, polyethylene glycols,glycerin, propylene glycol or other solvents; antibacterial agents;antioxidants; chelating agents; buffers such as acetates, citrates orphosphates and agents for the adjustment of tonicity such as sodiumchloride or dextrose. A parenteral preparation can be enclosed inampoules, disposable syringes or multiple dose vials made of glass orplastic. The use of physiological saline is preferred, and an injectablepharmaceutical composition is preferably sterile.

The methods of the present invention include application of SOSTantagonists in cocktails including other medicaments, for example,antibiotics, fungicides, and anti-inflammatory agents. Alternatively,the methods may comprise sequential dosing of an afflicted individualwith a SOST antagonist and one or more additional medicaments tooptimize a treatment regime. In such optimized regimes, the medicaments,including the granulation inhibitor can be applied in any sequence andin any combination.

The SOST, Wise, or LRP antagonists of the present invention may also beincluded in slow release formulations for prolonged treatment followinga single dose. In one embodiment, the formulation is prepared in theform of microspheres. The microspheres can be prepared as a homogenousmatrix of a SOST antagonist with a biodegradable controlled releasematerial, with optional additional medicaments as the treatmentrequires. The microspheres are preferably prepared in sizes suitable forinfiltration and/or injection, and injected systemically, or directly atthe site of treatment.

The formulations of the invention are also suitable for administrationin all body spaces/cavities, including but not limited to pleura,peritoneum, cranium, mediastinum, pericardium, bursae or bursal,epidural, intrathecal, intraocular, intra-articular, intra-discal,intra-medullary, perispinal, etc.

Some slow release embodiments include polymeric substances that arebiodegradable and/or dissolve slowly. Such polymeric substances includepolyvinylpyrrolidone, low- and medium-molecular-weight hydroxypropylcellulose and hydroxypropyl methylcellulose, cross-linked sodiumcarboxymethylcellulose, carboxymethyl starch, potassiummethacrylatedivinylbenzene copolymer, polyvinyl alcohols, starches,starch derivatives, microcrystalline cellulose, ethylcellulose,methylcellulose, and cellulose derivatives, β-cyclodextrin, poly(methylvinyl ethers/maleic anhydride), glucans, scierozlucans, mannans,xanthans, alzinic acid and derivatives thereof, dextrin derivatives,glyceryl monostearate, semisynthetic glycerides, glycerylpalmitostearate, glyceryl behenate, polyvinylpyrrolidone, gelatine,magnesium stearate, stearic acid, sodium stearate, talc, sodiumbenzoate, boric acid, and colloidal silica.

Slow release agents of the invention may also include adjuvants such asstarch, pregelled starch, calcium phosphate mannitol, lactose,saccharose, glucose, sorbitol, microcrystalline cellulose, gelatin,polyvinylpyrrolidone. methylcellulose, starch solution, ethylcellulose,arabic gum, tragacanth gum, magnesium stearate, stearic acid, colloidalsilica, glyceryl monostearate, hydrogenated castor oil, waxes, andmono-, bi-, and trisubstituted glycerides. Slow release agents may alsobe prepared as generally described in WO94/06416.

The amount of SOST, Wise, or LRP antagonists administered to anindividual will depend, in part, on the disease and/or extent of injury.Methods for determining an effective amount of an agent to administerfor a diagnostic or a therapeutic procedure are well known in the artand include phase I, phase II and phase III clinical trials, or thePilot and Pivotal trials (FDA device approval pathway). Generally, anagent antagonist is administered in a dose of about 0.01 to 200 mg/kgbody weight when administered systemically, and at a concentration ofapproximately 0.1-100 μM when administered directly to a wound site. Thetotal amount of SOST antagonist can be administered to a subject as asingle dose, either as a bolus or by infusion over a relatively shortperiod of time, or can be administered using a fractionated treatmentprotocol, in which the multiple doses are administered over a moreprolonged period of time. One skilled in the art would know that theconcentration of a particular SOST antagonist required to provide aneffective amount to a region or regions of injury depends on manyfactors including the age and general health of the subject as well asthe route of administration, the number of treatments to beadministered, and the nature of the SOST antagonist. In view of thesefactors, the skilled artisan would adjust the particular dose so as toobtain an effective amount for efficaciously promoting bone formationfor therapeutic purposes.

The compounds of the present invention can be formulated in a variety ofdifferent manners known to one of skill in the art. Pharmaceuticallyacceptable carriers are determined in part by the particular compositionbeing administered, as well as by the particular method used toadminister the composition. Accordingly, there are a wide variety ofsuitable formulations of pharmaceutical compositions of the presentinvention (see, e.g., Remington's Pharmaceutical Sciences, 20^(th) ed.,2003, supra).

Formulations suitable for oral administration can consist of (a) liquidsolutions, such as an effective amount of a compound of the presentinvention suspended in diluents, such as water, saline or PEG 400; (b)capsules, sachets, depots or tablets, each containing a predeterminedamount of the active ingredient, as liquids, solids, granules orgelatin; (c) suspensions in an appropriate liquid; (d) suitableemulsions; and (e) patches. The pharmaceutical forms can include one ormore of lactose, sucrose, mannitol, sorbitol, calcium phosphates, cornstarch, potato starch, microcrystalline cellulose, gelatin, colloidalsilicon dioxide, talc, magnesium stearate, stearic acid, and otherexcipients, colorants, fillers, binders, diluents, buffering agents,moistening agents, preservatives, flavoring agents, dyes, disintegratingagents, and pharmaceutically compatible carriers. Lozenge forms cancomprise the active ingredient in a flavor, e.g., sucrose, as well aspastilles comprising the active ingredient in an inert base, such asgelatin and glycerin or sucrose and acacia emulsions, gels, and the likecontaining, in addition to the active ingredient, carriers known in theart.

The pharmaceutical preparation is preferably in unit dosage form. Insuch form the preparation is subdivided into unit doses containingappropriate quantities of the active component. The unit dosage form canbe a packaged preparation, the package containing discrete quantities ofpreparation, such as packeted tablets, capsules, and powders in vials orampoules. Also, the unit dosage form can be a capsule, tablet, cachet,or lozenge itself, or it can be the appropriate number of any of thesein packaged form. The composition can, if desired, also contain othercompatible therapeutic agents. Preferred pharmaceutical preparations candeliver the compounds of the invention in a sustained releaseformulation.

The pharmaceutical preparations are typically delivered to a mammal,including humans and non-human mammals. Non-human mammals treated usingthe present methods include domesticated animals (i.e., canine, feline,murine, rodentia, and lagomorpha) and agricultural animals (bovine,equine, ovine, porcine).

In practicing the methods of the present invention, the pharmaceuticalcompositions can be used alone, or in combination with other therapeuticor diagnostic agents. The additional drugs used in the combinationprotocols of the present invention can be administered separately or oneor more of the drugs used in the combination protocols can beadministered together, such as in an admixture. Where one or more drugsare administered separately, the timing and schedule of administrationof each drug can vary. The other therapeutic or diagnostic agents can beadministered at the same time as the compounds of the present invention,separately or at different times.

V. Orthopedic and Periodontal Devices

In some embodiments, the present invention provides an orthopedic orperiodontal medical device formed from a structural support, wherein animplantable portion of the structural support is adapted to bepermanently implanted within a subject, wherein the implantable portionis attached to a bone, the structural support bearing at least a partialexternal coating including a compound of the present invention.

Other aspects of the present invention are directed towards medicalimplants. Such medical devices and implants include, for example, theosteogenic devices and methods of using the same for repairingendochondral bone and osteochondral defects taught in US patentapplication publication No. 20060177475 to David Rueger et al.,published Aug. 10, 2006, as well as in issued U.S. Pat. Nos. 6,190,880,5,344,654, 5,324,819, 5,468,845, 6,949,251, 6,426,332 and 5,656,593, andU.S. Publication Nos. 2002/0169122, 2002/0187104, 2006/0252724 and2007/0172479, the subject matter of which is hereby incorporated byreference.

These medical devices generally provide a structural support having animplantable portion preferentially adapted to mechanically engage boneand/or cartilage as taught, for instance, in U.S. Publication No.2006/0178752 to Joseph Vaccarino III, et al., published Aug. 10, 2006,the subject matter of which is hereby incorporated by reference. Thesebone implants desirably comprise an active agent on at least a portionthereof. As shown by U.S. Publication No. 2006/0188542 to John DennisBobyn, et al., published Aug. 24, 2006, the subject matter of which ishereby incorporated by reference, the active agent is preferablyformulated to be locally deliverable to bone proximate the implant insustained-release or in at least a two-phased release scheme. In thelatter, a first phase rapidly releases a first quantity of the activeagent, and the second and subsequent phases gradually release a secondquantity of the active agent, whereby bone formation stimulated by theactive agent is modulated.

Medical devices such as bone implants feature implantable portionsbearing Sost antagonists foster quicker and more complete bone formationin situ. The implantable portion of the medical device can be desirableat least partially or totally covered or impregnated with a Sostantagonist. In some embodiments, the external coating completely coatsthe implantable portion of the structural support.

In some other embodiments, the implantable portion of the structuralsupport comprises an osteoconductive matrix. The matrix material can beconducive to bone growth. This can be desirable for materials such asteeth and artificial bone graft sections, and the like. Alternatively,when the implantable sections are load bearing and formed, e.g., ofstainless steel, these implantable sections can be desirable when formedwith a Sost antagonist coating. In that event, it is desirable to alsoprovide a separate matrix material conducive to forming new bone growth.

Suitable matrixes include those comprising composite biomaterials havinga sponge-like structure such as those containing, e.g., phosphosphorynand/or collagen as taught in Takashi Saito's U.S. Publication No.2006/0188544, published Aug. 24, 2006, the subject matter of which ishereby incorporated by reference. Such coatings include, for example,the single and multilayer coatings taught in U.S. Publication No.2006/0204542 to Zongtao Zhang et al, published Sep. 14, 2006, as well asthose in U.S. Pat. Nos. 6,949,251, 5,298,852, 5,939,039, and 7,189,263and can be made by conventional methods including the methods taughttherein, the subject matter of which is hereby incorporated byreference.

The matrix can be part of the device of the present invention. In otherembodiments, the osteoconductive matrix includes an osteoinductive agentsuch as bone allograft, bone autograft, demineralized bone orperiodontal ligament cells. In some other embodiments, theosteoconductive matrix includes a calcium salt, calcium sulfate, calciumphosphate, a calcium phosphate cement, hydroxyapatite, coralline basedhydroyxapatite (HA), dicalcium phosphate, tricalcium phosphate (TCP),calcium carbonate, collagen, plaster of Paris, phosphosphoryn, aborosilicate, a biocompatible ceramic, a calcium phosphate ceramic andpolytetrafluoroethylene. One of skill in the art will appreciate thatother osteconductive matrices and osteoinductive agents are useful inthe present invention.

VI. Assay for Identification of Compounds for Treating Bone Loss

Compounds useful in the methods of the present invention can beidentified via a variety of methods known to one of skill in the art.Several exemplary methods for identifying such antagonists are describedherein, including cell-based and in vitro techniques (Journal of Boneand Mineral Research 2006, 21(11), 1738-1749). A general method ofidentifying SOST antagonists involves evaluating the effects ofantagonist candidates on bone formation under controlled conditions.Preferably bone formation is determined using micro-CT techniques onlive animals. Preferred animals include rodents, more preferred areprimates. Femur, tibia and vertebrae bones are particularly usefulsubjects for such study.

Briefly, the test animal is treated with a predetermined dose of a SOSTantagonist candidate. A control animal is treated with a controlsolution, preferably a non-irritating buffer solution or other carrier.

Once the dosing schedule has been completed, both test and controlanimals are examined to determine the quantity of bone formationpresent. This can be accomplished by any suitable method, but ispreferably performed on live animals to analyze the bone mineralcontent. Methods for micro-CT examination of bones in animals are wellknown in the art. A SOST antagonist candidate suitable for use as a SOSTantagonist is identified by noting significant bone formation in thetest animal when compared to the control animal. Bone formation in thetest bone(s) of the test animal can be 0.5%, 1, 5, 10, 20, 30, 40, 50,60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, 900 or 1000%more bone formation than is present in the same bones of the controlanimal. More preferably, bone formation can be 20%, most preferably 30%or 40%. Where necessary, levels of bone formation can be calculated bydetermining the volume of bone formation present in each animal.Calculations can be performed by constructing a 3-dimensional image ofthe bone formation and calculating the volume from the image with theaid of e.g., histomorphometry.

An example of the molecular modelling system described generally aboveconsists of the CHARMm and QUANTA programs, Polygen Corporation,Waltham, Mass. CHARMm performs the energy minimization and moleculardynamics functions. QUANTA performs the construction, graphic modellingand analysis of molecular structure. QUANTA allows interactiveconstruction, modification, visualization, and analysis of the behaviorof molecules with each other.

SOST antagonists may also be identified using a process known ascomputer, or molecular modeling, which allows visualization of thethree-dimensional atomic structure of a selected molecule and therational design of new compounds that will interact with the molecule.The three-dimensional construct typically depends on data from x-raycrystallographic analyses or NMR imaging of the selected molecule. Themolecular dynamics require force field data. The computer graphicssystems enable prediction of how a new compound will link to the targetmolecule and allow experimental manipulation of the structures of thecompound and target molecule to perfect binding specificity. Predictionof what the molecule-compound interaction will be when small changes aremade in one or both requires molecular mechanics software andcomputationally intensive computers, usually coupled with user-friendly,menu-driven interfaces between the molecular design program and theuser.

VII. Method of Promoting Bone Growth

In some embodiments, the present invention provides a method ofpromoting bone growth in a subject in need thereof, by administering tothe subject a therapeutically effective amount of a compound of thepresent invention.

Bone growth can be measured in a variety of ways known to one of skillin the art. Methods of measuring bone growth include, but are notlimited to, Uct (micro CT), Dual X-ray absorption (Bone density),ultrasound, QCT, SPA, DPA, DXR, SEXA, QUS, X-ray, using the human eyeduring surgically manipulation, Alizarin red S, serum osteocalcin, serumalkaline phosphatase, Serum bone Gla-protein (BGP), bone mineralcontent, serum calcium, serum phosphorus, tantalum markers, and serumIGF-1.

Many indicators of bone growth can be used to measure bone growth,including bone density. In some embodiments, bone growth can bedemonstrated by an increase of 0.1% in bone density. In otherembodiments, bone growth can be demonstrated by an increase of 0.2, 0.3,0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30,40, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, 900 or1000% or greater, in bone density.

One of skill in the are appreciates that bone growth be local, systemicor both.

A. Local Bone Growth

In some embodiments, the present invention provides a method ofpromoting bone growth at a site of injury or localized condition. Asubject in need of local bone growth can suffer from a variety ofailments and disease states. In other embodiments, the injury can be afracture or weakened bone. In some other embodiments, the subject can bein need of a spinal fusion, arthrodesis or an orthopedic or periodontalsynthetic bone graft or implant.

The local bone growth of the present invention can be achieved in avariety of methods. In some embodiments, the method further comprisesadministering to the subject a osteoconductive matrix, as describedabove. The matrix can be part of the device of the present invention, asdescribed above. In other embodiments, the osteoconductive matrixincludes an osteoinductive agent such as bone allograft, bone autograft,demineralized bone or periodontal ligament cells. In some otherembodiments, the osteoconductive matrix includes a calcium salt, calciumsulfate, calcium phosphate, a calcium phosphate cement, hydroxyapatite,coralline based hydroyxapatite (HA), dicalcium phosphate, tricalciumphosphate (TCP), calcium carbonate, collagen, plaster of Paris,phosphosphoryn, a borosilicate, a biocompatible ceramic, a calciumphosphate ceramic and polytetrafluoroethylene.

B. Systemic Bone Growth

In other embodiments, the present invention provides a method ofpromoting systemic bone growth. Systemic bone growth refers to thegrowth of bone throughout the subject, and can effect all the bones inthe subject's body. A subject in need of systemic bone growth can sufferfrom a variety of ailments and disease states. In some embodiments, thesubject suffers from a low bone mass phenotype disease. Low bone masscan be determined by a variety of methods known to one of skill in theart. For example, low bone mass can be characterized by a T-score lessthan about −1. Low bone mass phenotype diseases can includeosteoporosis, osteopenia, and osteoporosis-pseudoglioma syndrome (OPPG).In some other embodiments, the low bone mass phenotype disease can beosteopenia or osteoporosis-pseudoglioma syndrome (OPPG).

The methods of the present invention can also be used to treat diseasescharacterized by secondary induced osteoporosis (low bone mass)including, but not limited to, osteomalacia, Polyostotic fibrousdysplasia, Paget's disease, rheumatoid arthritis, zero gravity,osteoarthritis, Prolonged inactivity or immobility, osteomyelitis,Celiac disease, Crohn's Disease, Ulcerative Colitis, inflammatory bowldisease, gastrectomy, secondary induced osteoporosis, Amennorhea,Cushing's Disease, Cushing's syndrome, Diabetes Mellitus, Diabetes,Eating Disorders, Hyperparathyroidism, Hyperthyroidism,Hyperprolactinemia, Kleinefelter Syndrome, Thyroid Disease, TurnerSyndrome, steroid induced osteoporosis, seizure or depression inducedosteoporosis, immobility, arthritis, cancer induced secondaryosteoporosis, Gonadotropin-releasing hormone agonists induced low bonemass, Thyroid medication induced low bone mass, Dilantin (phenytoin),depakote induced low bone mass, chemotherapy induced low bone mass,Immunosuppressant induced low bone mass, Blood thinning agents inducedlow bone mass, Grave's disease, Juvenile rheumatoid arthritis,Malabsorption syndromes, Anorexia nervosa, Kidney disease,Anticonvulsant treatment (e.g., for epilepsy), Corticosteroid treatment(e.g., for rheumatoid arthritis, asthma), Immunosuppressive treatment(e.g., for cancer), Inadequate nutrition (especially calcium, vitaminD), Excessive exercise leading to amenorrhea (absence of periods),Smoking, and Alcohol abuse, pregnancy-associated osteoporosis, copperdeficiency, Dibasic aminoaciduria type 2, Werner's syndrome,Hajdu-Cheney syndrome, Hyperostosis corticalis deformans juvenilis,Methylmalonic aciduria type 2, Cystathionine beta-synthase deficiency,Exemestane, Hyperimmunoglobulin E (IgE) syndrome, Haemochromatosis,Singleton-Merten syndrome, Beta thalassaemia (homozygous), Reflexsympathetic osteodystrophy, Sarcoidosis, Winchester syndrome,Hallermann-Streiff syndrome (HSS), Cyproterone, Glycerol kinasedeficiency, Bonnet-Dechaume-Blanc syndrome, Prednisolone, Heparin,Geroderma osteodysplastica, Torg osteolysis syndrome, Orchidectomy,Fabry's disease, Pseudoprogeria syndrome, Wolcott-Rallison syndrome,Ankylosing spondylitis, Myeloma, Systemic infantile hyalinosis,Albright's hereditary osteodystrophy, Anorexia Nervosa, AutoimmuneLymphoproliferative Syndrome, Brown-Sequard Syndrome, Diamond-Blackfananemia, Eating disorders, Galactorrhoea-Hyperprolactinaemia, Gonadaldysgenesis, Kidney conditions, Menkes Disease, Menopause, Neuritis,Ovarian insufficiency due to FSH resistance, Familial Ovarianinsufficiency, Premature aging, Primary biliary cirrhosis, Prolactinoma,Familial Prolactinoma, Renal osteodystrophy, Ulcerative colitis,Underweight, Werner syndrome, Bone tumor, Bone cancer, Brittle bonedisease, Osteogenesis imperfecta congenita, and Osteogenesis imperfectatarda. One of skill in the art will appreciate that other types ofconditions, diseases and treatments lead to osteoporosis.

Following administration of the compounds of the present invention,systemic bone growth can be determined by a variety of methods, such asimprovements in bone density. Bone density can be measured by a varietyof different methods, including the T-score and Z-score. The Z-score isthe number of standard deviations above or below the mean for thepatient's age and sex. The T-score is the number of standard deviationsabove or below the mean for a healthy 30 year old adult of the same sexas the patient. Low bone mass is characterized by a T-score of −1 to−2.15. Osteoporosis is characterized by a T-score less than −2.15.Improvement in the T-score or Z-score indicate bone growth. Bone densitycan be measured in a variety of places of the skeleton, such the spineor the hip. One of skill in the art will appreciate that other methodsof determining bone density are useful in the present invention.

C. Promoting Bone Growth with a Compound of the Present Invention and anAntiresorptive Drug

In some other embodiments, the method of the present invention promotesbone growth by administering the compound of Formula I with anantiresorptive drug. Antiresorptive drugs include those that slow orblock the resorption of bone. Administration of a compound of Formula Iand an antiresorptive drug can promote local bone growth and/or systemicbone growth. In some embodiments, the administration of a compound ofFormula I and an antiresorptive drug promotes systemic bone growth. Bonegrowth can be achieved by increasing bone mineral content, increasingbone density and/or growth of new bone. In other embodiments, localapplication of the compound of Formula I and an antiresorptive drugachieves systemic bone growth.

Antiresorptive drugs useful in the methods of the present inventioninclude, but are not limited to, denosumab, a RankL inhibitor, abisphosphonate, a selective estrogen receptor modulator (SERM),calcitonin, a calcitonin analog, Vitamin D and a Vitamin D analog.

In other embodiments, the antiresorptive drug can be a bisphosphonate(i.e. fosamax, actonel, reclast), a parathyroid hormone (PTH) or analog(i.e. teriparatide (Forteo)), calcitonin or analog (i.e. Miacalcic),Vitamin D or analog, SERM or analog (i.e. Evista).

Bisphosphonates useful in the methods of the present invention can beany suitable bisphosphonate. In some embodiments, the bisphosphonatesare nitrogenous, such as Pamidronate (APD, Aredia), Neridronate,Olpadronate, Alendronate (Fosamax), Ibandronate (Boniva), Risedronate(Actonel) and Zoledronate (Zometa). In other embodiments, thebisphosphonates are non-nitrogenous, such as Etidronate (Didronel),Clodronate (Bonefos, Loron) and Tiludronate (Skelid). One of skill inthe art will appreciate that other bisphosphonates are useful in thepresent invention.

SERMs useful in the methods of the present invention can be any suitableSERM. In some embodiments, the SERM can be clomifene, raloxifene,tamoxifen, toremifene, bazedoxifene, lasofoxifene or ormeloxifene. Oneof skill in the art will appreciate that other SERMs are useful in thepresent invention.

The antiresorptive drug can also be any suitable calcitonin analog. Insome embodiments, calcitonin analogs useful in the methods of thepresent invention include, but are not limited to, miacalcic. One ofskill in the art will appreciate that other calcitonin analogs areuseful in the present invention.

Vitamin D analogs useful in the methods of the present invention can beany suitable Vitamin D analog. In some embodiments, Vitamin D analog'suseful in the methods of the present invention include, but are notlimited to, Vitamin D1 (molecular compound of ergocalciferol withlumisterol, 1:1), Vitamin D2 (ergocalciferol or calciferol), Vitamin D3(cholecalciferol), Vitamin D4 (22-dihydroergocalciferol) and Vitamin D5(sitocalciferol). One of skill in the art will appreciate that otherVitamin D analogs are useful in the present invention.

RankL inhibitors useful in the present invention include any compoundsthat inhibit the activity of RankL. For example, RankL inhibitorsinclude, but are not limited to, the human monoclonal antibodydenosumab. One of skill in the art will appreciate that other RankLinhibitors are useful in the present invention.

VIII. Treating Renal Damage

In some embodiments, the present invention provides a method of treatingrenal damage by administering to a subject suffering from renal damage,a therapeutically effective amount of a compound of Formula I.

Renal damage can be caused by a variety of ailments known to one ofskill in the art. In some embodiments, renal damage is caused byinfection, radiation, toxin, dehydration or trauma. Toxins causing renaldamage include, but are not limited to, chemicals, poisons, andchemotherapeutic agents. One of skill in the art will appreciate thatother causes of renal damage can be treated by the methods of thepresent invention.

Renal damage treatable by the compounds of the present inventionincludes acute renal failure. Acute renal failure is also known as acutekidney failure or acute kidney injury. Acute renal failure results inretention of nitrogenous (urea and creatinine) and non-nitrogenous wasteproducts that are normally excreted by the kidney. Depending on theseverity and duration of the renal dysfunction, this accumulation isaccompanied by metabolic disturbances, such as metabolic acidosis(acidification of the blood) and hyperkalaemia (elevated potassiumlevels), changes in body fluid balance, and effects on other organsystems. Acute renal failure can be characterized by oliguria or anuria(decrease or cessation of urine production), although nonliguric acuterenal failure can also occur.

A subject can be characterized as being at (1) a risk for acute damage;(2) kidney damage resulting in injury; (3) acute renal failure; and (4)loss of kidney function. Risk for acute kidney damage is characterizedby serum creatinine increased 1.5 times or urine production of <0.5ml/kg body weight over 6 hours. Injury is reached when serum creatinineincreased 2.0 times or urine production<0.5 ml/kg over 12 hours. Failureis reached when serum creatinine increased 3.0 times or creatinine>355μM (with a rise of >44) or urine output below 0.3 ml/kg over 24 hours.Loss of kidney function is reached when a subject suffers frompersistent acute renal failure or more than four weeks of complete lossof kidney function.

Kidney biopsy can be performed in the setting of acute renal failure, toprovide a definitive diagnosis and sometimes an idea of the prognosis,unless the cause is clear and appropriate screening investigations arereassuringly negative.

Renal therapeutic agents of the invention can be used in subjects thathave received renal injury, or those at risk of chronic renal failure.As used herein, a subject is said to be in, or at risk or, chronic renalfailure, or at risk of the need for renal replacement therapy (i.e.,chronic hemodialysis, continuous peritoneal dialysis, or kidneytransplantation), if the subject is reasonably expected to suffer aprogressive loss of renal function associated with progressive loss offunctioning nephron units. Whether a particular subject is in, or atrisk of, chronic renal failure is a determination which may routinely bemade by one of ordinary skill in the relevant medical or veterinary art.Subjects in, or at risk of, chronic renal failure, or at risk of theneed for renal replacement therapy, include but are not limited to thefollowing: subjects which can be regarded as afflicted with chronicrenal failure, end-stage renal disease, chronic diabetic nephropathy,hypertensive nephrosclerosis, chronic glomerulonephritis, hereditarynephritis, and/or renal dysplasia; subjects having a biopsy indicatingglomerular hypertrophy, tubular hypertrophy, chronic glomerulosclerosis,renal cell carcinoma, and/or chronic tubulointerstitial sclerosis;subjects having an ultrasound, MRI, CAT scan, or other non-invasiveexamination indicating renal fibrosis; subjects having an unusual numberof broad casts present in urinary sediment; subjects having a GFR whichis chronically less than about 50%, and more particularly less thanabout 40%, 30% or 20%, of the expected GFR for the subject; human malesubjects weighing at least about 50 kg and having a GFR which ischronically less than about 50 ml/min, and more particularly less thanabout 40 ml/min 30 ml/min or 20 ml/min; human female subjects weighingat least about 40 kg and having a GFR which is chronically less thanabout 40 ml/min, and more particularly less than about 30 ml/min, 20ml/min or 10 ml/min; subjects possessing a number of functional nephronunits which is less than about 50%, and more particularly less thanabout 40%, 30% or 20%, of the number of functional nephron unitspossessed by a healthy but otherwise similar subject; subjects whichhave a single kidney; and subjects which are kidney transplantrecipients.

IX. Treating Cancer

The compounds and compositions of the present invention are also usefulin the treatment of cancer. The compounds of formula I can possessanti-proliferative activity and are therefore useful in the treatment ofproliferative disorders such as cancers, leukaemias and other disordersassociated with uncontrolled cellular proliferation such as psoriasisand restenosis. As defined herein, an anti-proliferative effect withinthe scope of the present invention may be demonstrated by the ability toinhibit cell proliferation in an in vitro whole cell assay, for exampleusing any of the cell lines A549, HT29, Saos-2, HeLa or MCF-7, or byshowing inhibition of a CDK enzyme (such as CDK2 or CDK4) in anappropriate assay. Using such cell line and enzymes assays it may bedetermined whether a compound is anti-proliferative in the context ofthe present invention.

As used herein, the term “cancer” includes, but is not limited to thefollowing cancers: breast, ovary, cervix, prostate, testis,genitourinary tract, esophagus, larynx, glioblastoma, neuroblastoma,stomach, skin, keratoacanthoma, lung, epidermoid carcinoma, large cellcarcinoma, small cell carcinoma, lung adenocarcinoma, bone, colon,adenoma, pancreas, adenocarcinoma, thyroid, follicular carcinoma,undifferentiated carcinoma, papillary carcinoma, seminoma, melanoma,sarcoma, bladder carcinoma, liver carcinoma and biliary passages, kidneycarcinoma, myeloid disorders, lymphoid disorders, Hodgkin's, hairycells, buccal cavity and pharynx (oral), lip, tongue, mouth, pharynx,small intestine, colon-rectum, large intestine, rectum, brain andcentral nervous system, and leukemia. One of skill in the art willappreciate that other cancers and proliferative disorders can be treatedby the compounds and compositions of the present invention.

In some embodiments, the cancer is bone cancer, colon cancer, multiplemyeloma, gastric cancer, colorectal cancer, prostate cancer, cervicalcancer, lung cancer, pancreatic cancer, medulloblastoma, liver cancer,parathyroid cancer, endometrial cancer, or breast cancer. In otherembodiments, the cancer is bone cancer.

X. Examples Example 1 Promotion of Bone Growth

Using the assay described above and in Journal of Bone and MineralResearch 2006, 21(11), 1738-1749 (incorporated herein in its entirety),compounds of the present invention can be identified as promoting bonegrowth. For example, the mouse test animal is treated with apredetermined dose of a SOST antagonist candidate for a complete dosingschedule. A control mouse is treated with a control solution, preferablya non-irritating buffer solution or other carrier. Once the dosingschedule has been completed, both test and control animals are examinedwith sacrifice using micro-CT to determine the quantity of boneformation present.

Example 2 Bone Growth with Eburnamonine

Four month old male C57BL/6 mice were treated daily with saline vehicleor the sclerostin inhibitor of eburnamonine phosphate at several doses(via i.p) for 30 days. Study endpoints included a biochemical marker ofbone formation; osteocalcin, measured by EIA Elisa, and measurement oftrabecular bone volume by mico-CT analysis on the proximal tibia andspine (lumbar 5). FIG. 1 shows the increase in bone volume for thetibia, as measured by micro-CT: 17% at 5 mg/kg (B) and 10% at 1 mg/kg(C) of ebumamonine phosphate in mice, as measured by U-CT analysis. TheU-CT bone volume measurement of lumbar 5 showed a 52% increase in bonevolume. See also FIG. 3, showing increase in bone volume for the tibia:5 mg/kg (D) and 0.33 mg/kg (E). FIG. 2 and FIG. 4 shows the percentincrease in the bone formation marker osteocalcin at 5 mg/kg (FIG. 2)and 0.33 mg/kg (FIG. 4) of ebumamonine phosphate in mice.

Example 3 Bone Growth with Vinpocetine

Using the procedure set forth in Example 2, vinpocetine citrate wasinvestigated for bone growth. Measurement of the tibia via pQCT showedan 8% increase in bone volume following IP treatment with vinpocetinecitrate at 10 mg/kg. FIG. 4 shows the increase in serum osteocalcin overbaseline for treatment with vinpocetine citrate at 10 mg/kg.

Although the foregoing invention has been described in some detail byway of illustration and example for purposes of clarity ofunderstanding, one of skill in the art will appreciate that certainchanges and modifications can be practiced within the scope of theappended claims. In addition, each reference provided herein isincorporated by reference in its entirety for all purposes to the sameextent as if each reference was individually incorporated by reference.

1. A method of promoting bone growth in a subject in need thereof,comprising administering to the subject a therapeutically effectiveamount of a compound of Formula I:

wherein each of R¹, R², R⁵, R⁷ and R⁸ are independently selected fromthe group consisting of H, halogen, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆haloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₁₋₆ haloalkoxy, C₁₋₆alkyl-hydroxy, —OR^(1a), —C(O)R^(1a), —C(O)OR^(1a), —C(O)NR^(1a)R^(1b),—NR^(1a)R^(1b), —SR^(1a), —N(R^(1a))C(O)R^(1b), —N(R^(1a))C(O)OR^(1b)—N(R^(1a))C(O)NR^(1a)R^(1b), —OP(O)(OR^(1a))₂, —S(O)₂OR^(1a),—S(O)₂NR^(1a)R^(1b), —CN, —CH═N—OH, C₁₋₆ alkyl-OC(O)-aryl, cycloalkyl,heterocycloalkyl, aryl and heteroaryl; alternatively, R¹ is combinedwith R² to form ═O or ═NR^(1a), R¹ is combined with R³ to form a bond,or R¹ is combined with R⁵ to form a cycloalkyl; each R^(1a) and R^(1b)is independently selected from the group consisting of H, C₁₋₆ alkyl,C₁₋₆ alkyl-hydroxy, C₁₋₆ alkyl-NO₂, C₁₋₆ alkyl-cycloalkyl,heterocycloalkyl, and heteroaryl; each of R³ and R⁴ are independentlyselected from the group consisting of H, C₁₋₆ alkyl and C₁₋₆ haloalkyl;alternatively two R⁵ or R⁷ groups attached to the same carbon can becombined to form ═O; R⁶ is absent or an N-oxide; and salts, hydrates andisomers thereof, thereby promoting bone growth in the subject.
 2. Themethod of claim 1, wherein the compound has the formula:


3. The method of claim 2, wherein the compound has the formula:


4. The method of claim 3, wherein R^(1a) is C₁₋₆ alkyl; and R⁴ is C₁₋₆alkyl.
 5. The method of claim 4, wherein the compound has the formula:


6. The method of claim 1, wherein the compound has the formula:


7. The method of claim 6, wherein the compound has the formula:


8. The method of claim 1, wherein the bone growth is promoted at a siteof injury or localized condition.
 9. The method of claim 8, wherein thebone growth is promoted at a site selected from the group consisting ofa bone fracture and weakened bone.
 10. The method of claim 8, whereinthe subject requires a spinal fusion, arthrodesis or an orthopedic orperiodontal synthetic bone graft or implant.
 11. The method of claim 8,further comprising the step of administering to the subject anosteoconductive matrix.
 12. The method of claim 11, wherein theosteoconductive matrix comprises an osteoinductive agent selected fromthe group consisting of bone allograft, bone autograft, demineralizedbone and periodontal ligament cells.
 13. The method of claim 11, whereinthe osteoconductive matrix comprises a calcium salt, calcium sulfate,calcium phosphate, a calcium phosphate cement, hydroxyapatite, corallinebased hydroyxapatite (HA), dicalcium phosphate, tricalcium phosphate(TCP), calcium carbonate, collagen, plaster of Paris, phosphosphoryn, aborosilicate, a biocompatible ceramic, a calcium phosphate ceramic andpolytetrafluoroethylene.
 14. The method of claim 1, wherein the bonegrowth is systemic.
 15. The method of claim 14, wherein the subjectsuffers from a low bone mass phenotype disease.
 16. The method of claim15, wherein the low bone mass phenotype disease is selected from thegroup consisting of osteoporosis, osteopenia, andosteoporosis-pseudoglioma syndrome (OPPG).
 17. The method of claim 1,wherein the compound is administered in combination with anantiresorptive drug.
 18. The method of claim 17, wherein theantiresorptive drug is selected from the group consisting of denosumab,a RankL inhibitor, a bisphosphonate, a selective estrogen receptormodulator (SERM), calcitonin, a calcitonin analog, Vitamin D and aVitamin D analog.
 19. The method of claim 17, wherein the bone growth issystemic.
 20. The method of claim 17, wherein the bone growth ispromoted by a local application of the compound and the antiresorptivedrug.
 21. A method of treating renal damage, comprising administering toa subject in need thereof, a therapeutically effective amount of acompound of Formula I:

wherein each of R¹, R², R⁵, R⁷ and R⁸ are independently selected fromthe group consisting of H, halogen, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆haloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₁₋₆ haloalkoxy, C₁₋₆alkyl-hydroxy, —OR^(1a), —C(O)R^(1a), —C(O)OR^(1a), —C(O)NR^(1a)R^(1b),—NR^(1a)R^(1b), —SR^(1a), —N(R^(1a))C(O)R^(1b), —N(R^(1a))C(O)OR^(1b),—N(R^(1a))C(O)NR^(1a)R^(1b), —OP(O)(OR^(1a))₂, —S(O)₂OR^(1a),—S(O)₂NR^(1a)R^(1b), —CN, —CH═N—OH, C₁₋₆ alkyl-OC(O)-aryl, cycloalkyl,heterocycloalkyl, aryl and heteroaryl; alternatively, R¹ is combinedwith R² to form ═O or ═NR^(1a), R¹ is combined with R³ to form a bond,or R¹ is combined with R⁵ to form a cycloalkyl; each R^(1a) and R^(1b)is independently selected from the group consisting of H, C₁₋₆ alkyl,C₁₋₆ alkyl-hydroxy, C₁₋₆ alkyl-NO₂, C₁₋₆ alkyl-cycloalkyl,heterocycloalkyl, and heteroaryl; each of R³ and R⁴ are independentlyselected from the group consisting of H, C₁₋₆ alkyl and C₁₋₆ haloalkyl;alternatively two R⁵ or R⁷ groups attached to the same carbon can becombined to form ═O; R⁶ is absent or an N-oxide; and salts, hydrates andisomers thereof, thereby treating renal damage in the subject.
 22. Anorthopedic or periodontal medical device comprising a structuralsupport, wherein an implantable portion of the structural support isadapted to be permanently implanted within a subject, wherein theimplantable portion is attached to a bone, the structural supportbearing at least a partial external coating comprising a compound ofclaim
 1. 23. A method of treating cancer, comprising administering to asubject in need thereof, a therapeutically effective amount of acompound of Formula I:

wherein each of R¹, R², R⁵, R⁷ and R⁸ are independently selected fromthe group consisting of H, halogen, C₁₋₆ alkyl, C₁₋₆ alkoxy, C₁₋₆haloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₁₋₆ haloalkoxy, C₁₋₆alkyl-hydroxy, —OR^(1a), —C(O)R^(1a), —C(O)OR^(1a), —C(O)NR^(1a)R^(1b),—NR^(1a)R^(1b), —SR^(1a), —N(R^(1a))C(O)R^(1b), —N(R^(1a))C(O)OR^(1b),—N(R^(1a))C(O)NR^(1a)R^(1b), —OP(O)(OR^(1a))₂, —S(O)₂OR^(1a),—S(O)₂NR^(1a)R^(1b), —CN, —CH═N—OH, C₁₋₆ alkyl-OC(O)-aryl, cycloalkyl,heterocycloalkyl, aryl and heteroaryl; alternatively, R¹ is combinedwith R² to form ═O or ═NR^(1a), R¹ is combined with R³ to form a bond,or R¹ is combined with R⁵ to form a cycloalkyl; each R^(1a) and R^(1b)is independently selected from the group consisting of H, C₁₋₆ alkyl,C₁₋₆ alkyl-hydroxy, C₁₋₆ alkyl-NO₂, C₁₋₆ alkyl-cycloalkyl,heterocycloalkyl, and heteroaryl; each of R³ and R⁴ are independentlyselected from the group consisting of H, C₁₋₆ alkyl and C₁₋₆ haloalkyl;alternatively two R⁵ or R⁷ groups attached to the same carbon can becombined to form ═O; R⁶ is absent or an N-oxide; and salts, hydrates andisomers thereof, thereby promoting bone growth in the subject.
 24. Themethod of claim 23, wherein the cancer is bone cancer, colon cancer,multiple myeloma, gastric cancer, colorectal cancer, prostate cancer,cervical cancer, lung cancer, pancreatic cancer, medulloblastoma, livercancer, parathyroid cancer, endometrial cancer, and breast cancer.